Combined preparation for the treatment of neoplasic diseases or of infectious diseases

ABSTRACT

The present invention relates to combined preparation containing as active substance the following individual components, in the form of a kit-of-parts: monocyte derived cells, particularly cytotoxic macrophages, chemotherapy or immunotherapy drugs, for the simultaneous, separate or sequential use, for the treatment of cancer or infectious diseases.

This is a 371 of PCT/EP99/02105, filed Mar. 29, 1999.

The present invention relates to a new combined preparation for thetreatment of neoplasic diseases or of infectious diseases.

The present invention describes sequences of conventional treatments ofcancer or infections and of immunotherapies reversing or preventingchemoresistance and allowing long lasting therapeutic responses.

BACKGROUND OF THE INVENTION

In conventional therapy, residual tumor cells or infectious agents areleft undamaged due to chemoresistance or due to the fact that thesecells are shaded in protected areas or located in hypoxic areas poorlyvascularized and not accessible to conventional treatments. The geneticinstability and heterogeneity of tumors and micro-organisms indeed allowthem to adapt and to develop resistance to therapies.

The beneficial effects of chemotherapy can be compromised by cellularmechanisms that allow infectious agents or neoplasic tissue to evade thetoxicity of drugs. In some cases, pleiotropic resistance to a variety ofunrelated drugs has been observed, and this phenomenon has been calledmultidrug resistance.

Resistance to chemotherapy, whether it is intrinsic or acquired, is amajor cause of failure in the curative treatment of chronic infectionsor neoplasic malignancies. Among the most active anti-cancer agents usedin the treatment of haematological malignancies are some naturaltoxin-derived drugs, such as the anthracycline daunorubicin oradriamicin, the epipodophyllotoxins, taxoter derivatives, the vincaalkaloid vincristine, cisplatin, fluorouracils.

Development of cross-resistance to these structurally and functionallyunrelated drugs, called multidrug resistance, is frequently observed insecond or third intention cytotoxic treatment of cancer.

Multiple drug resistance of infectious agents and particularly ofbacteria to antibiotics such as penicillins, β-lactamines,cephalosporines, aminoglucosides, macrolides and sulfamides, is more andmore often seen in hospitals.

Monocyte derived cells (MDCs) are immune cells such as obtained byculture of blood mononuclear cells in non adherent gas permeable plasticor Teflon bags for 5 to 10 days at 37° C. in O₂/CO₂ atmosphere. Theirculture medium (RPMI, IMDM, AIM5 (Gibco) or X-VIVO (Biowhittaker))contains eventually cytokines or ligands as defined in patentsPCT/EP93/01232, WO94/26875 or EP 97/02703 or in the articles mentionedbelow:

“Autologous lymphocytes prevent the death of monocytes in culture andpromote, as do GM-CSF, IL-3 and M-CSF, their differentiation intomacrophages”. (Lopez M., Martinache Ch., Canepa S., Chokri M., ScottoF., Bartholeyns J.; J. of Immunological Methods, 159: 29-38, 1993);

“Immune therapy with macrophages: Present status and criticalrequirements for implementation” (Bartholeyns J., Romet-Lemonne J-L.,Chokri M., Lopez M.; Immunobiol., 195: 550-562, 1996);

“In vitro generation of CD83⁺ human blood dendritic cells for activetumor immunotherapy” (Thurnher M., Papesh C., Ramoner R., Gastlt G. andal.; Experimental Hematology, 25: 232-237, 1997);

“Dendritic cells as adjuvants for immune-mediated resistance to tumors”(Schuler G. and Steinman R. M.; J. Exp. Med., 186: 1183-1187, 1997).

All these patents applications and articles are included herein forreferences.

They can be activated by IFN-γ at the end of culture to obtain inparticular cytotoxic macrophages. They can be centrifuged to beconcentrated and purified before resuspension in isotonic solution.

Monocyte derived cells (MDCs) can either be killer macrophages,phagocytozing cells, growth factors and cytokines releasing cells, ordendritic cells according to their conditions of differentiation.Dendritic cells can for example be obtained as described in “In vitrogeneration of CD83⁺ human blood dendritic cells for active tumorimmunotherapy” (Thurnher M., Papesh C., Ramoner R., Gastlt G. and al.;Experimental Hematology, 25: 232-237, 1997) and “Dendritic cells asadjuvants for immune-mediated resistance to tumors” (Schuler G. andSteinman R. M.; J. Exp. Med., 186: 1183-1187, 1997), and EP 97/02703.

In addition, activated monocyte derived cells (macrophages) can be usedto deliver therapeutic agents to tumor or infectious sites.

BRIEF SUMMARY OF THE INVENTION

One of the aims of the invention is to provide a combined preparation ofactive substances under the form of individual components for thesimultaneous separate or sequential use, in the treatment of cancer orof infectious diseases.

Another aim of the invention is to provide a method for the treatment ofresidual cancer resistant to chemotherapy.

Another aim of the invention is to provide a method for the treatment ofinfectious diseases resistant to antibiotic treatment.

The invention relates to a combined preparation containing, as activesubstance, the following individual components, in the form of akit-of-parts:

monocyte derived cells, particularly cytotoxic macrophages,

chemotherapy or immunotherapy drugs,

for the simultaneous, separate or sequential use, for the treatment ofcancer or infectious diseases.

BRIEF DESCRIPTION OF THE FIGURES

The present treatment consists in the local or systemic injection ofautologous activated macrophages (MAK® killer cells) or monocyte derivedcells which have access to injured areas, and in particular to hypoxicareas, where they tend to concentrate.

This treatment can be conducted after first failure and relapsefollowing chemotherapies, or before chemotherapy, to preventchemoresistance. Local treatment with chemotherapy drugs causes cellnecrosis and release of chemokines which call and actively recruitmacrophages and monocyte derived cells. Therefore, combining thechemotherapy with macrophage immunotherapy can in synergy increasecytotoxicity and increase immune response at the same time as preventingthe establishment of resistance. Additionally to a first treatmentcombining conventional approach with immunotherapy, macrophage adoptivetherapy can be proposed after failure and relapse.

It is shown through the invention that the local or systemic injectionof activated monocyte derived cells, or macrophages, restores clinicalresponses to cytotoxic drugs for which resistance was previouslydemonstrated, or prevents the apparition of chemoresistance.

The present invention also shows that activated monocyte derived cellscan overcome this resistance and synergize for therapy.

The two active ingredients of the combined preparation have never beenused for a new joint effect and are unknown as a composition.

The active ingredients which are administered either at the same time,or separately, or sequentially, according to the invention, do notrepresent a mere aggregate of known agents, but a new combination withthe surprising valuable property that immunotherapy with monocytederived cells modifies the chemoresistance/chemosensitivity and allows anew effective treatment (partial or complete response) with similarchemotherapy protocol. Furthermore, synergy is observed between monocytederived cells immunotherapy and chemotherapy.

It is to be stressed that the combined preparation also designated by akit-of-parts means that the components of the combined preparation arenot necessarily present as a union e.g. in composition, in order to beavailable for separate or sequential application. Thus the expressionkit-of-parts means that it is not necessarily a true combination, inview of the physical separation of the components.

In an advantageous combined preparation of the invention, the monocytederived cells contain chemotherapy or immunotherapy drugs.

In another advantageous combined preparation of the invention, themonocyte derived cells are such as prepared according to the methodcomprising the following steps:

1) recovery of blood derived mononuclear cells directly from bloodapheresis or from blood bag collection, followed if necessary bycentrifugation, to eliminate a substantial part of red blood cells,granulocytes and platelets, and collection of peripheral bloodleukocytes;

2) washing peripheral blood leukocytes obtained at the preceeding stepsfor instance by centrifugation (to remove 90% of platelets, red bloodcells and debris) to obtain mononuclear cells;

3) resuspension of the total mononuclear cells (monocytes+lymphocytes)obtained at the preceeding step in culture medium (RPMI or IMDM type) at10⁶ to 2.10⁷ cells/ml, possibly completed by cytokines and/or autologousserum, and culture for 5 to 10 days at 37° C. under O₂/CO₂ atmosphere inhydrophobic gas permeable bags, to obtain monocyte derived cells andcontaminating lymphocytes .

According to an advantageous combined preparation, the chemotherapy drugis selected among cytotoxic compounds such as anthracyclins,daunorubicin, adriamycin, taxoter derivatives, vinca alcaloids,vincristine, carmustine, cisplatin, fluorouracils, cytostatic compoundssuch as polyamine inhibitors, topoisomerase inhibitors, tamoxifene,prodasone, or sandostatine, or compounds inducing apoptosis such assodium butyrate or mitomycin C, antibiotics such as penicilins,β-lactamines, cephalosporines, cyclines, arninoglucosides, macrolides orsulfamides, or antiviral drugs such as AZT, protease inhibitors oracyclovir, retrovir or foscarnet.

According to an advantageous embodiment, the combined preparation of theinvention is such that the immunotherapy drug is selected amongcytokines such as cyclosporine, azathioprine, cyclophosphamide, IFNγ,IL-12, IL-2, GM-CSF, G-CSF, adjuvants such as murapeptides or BCG, orvaccines with or without substances with adjuvant effect. The vaccinescan be constituted by tumor or infectious antigens which are of natural,recombinant or gene transfer origin, formulated in the presence or notof an adjuvant for the administration to humans. The vaccine can also bea nucleic acid coding for the antigen or a fragment of antigen, or avirus expressing the antigen.

According to an advantageous embodiment, in the combined preparation ofthe invention, the monocyte derived cells and the chemotherapy orimmnunotherapy drugs are in the form in injectable solutions.

In another advantageous embodiment of the invention, in the combinedpreparation, the injectable solutions are in the form of locallyinjectable solutions.

In another advantageous embodiment in the combined preparation of theinvention, the injectable solutions are in the form of systemicallyinjectable solutions.

In another advantageous combined preparation of the invention, themonocyte derived cells are administered at a dose of about 10⁷ to about10¹⁰ monocyte derived cells per injection.

In another advantageous combined preparation of the invention, themonocyte derived cells are administrated at a dose of about 10⁸ to about10⁹.

In another advantageous combined preparation of the invention, themonocyte derived cells are administered in a repeated way up to tentimes, the interval between each administration being between three daysto two months.

In another advantageous combined preparation of the invention, theimmunotherapy or chemotherapy drug is administered at a dose of about0.1 to about 1000 mg/day.

In another advantageous combined preparation of the invention, in thecase of administration of a drug chosen among immunotherapy drug,antiviral drug, cytotoxic drugs, or antibiotics, said drug isadministered at a dose of about 10 to about 1000 mg/day.

More specifically, in the case of cytotoxic compounds such asvincristine, taxol, carmustine, daunorubicin, adryamicin, cisplatin,fluorouracil, they are administered at a dose of about 10 to about 500mg/day.

In the case of antiviral drugs such as retrovir, aciclovir, foscarnet,said drug is administered at a dose of about 20 to about 500 mg/day.

In the case of antibiotics such as penicilins, cephalosporine,sulfamides, cyclines, said drug is administered at a dose of about 10 toabout 1000 mg/day.

In the case of immunotherapy drugs such as cyclosporine, azathioprine,cyclophosphamide, said drug is administered at a dose of about 10 toabout 1000 mg/day.

In another advantageous combined preparation of the invention, in thecase of administration of a drug chosen among cytostatic compounds,apoptosis inducing compounds or cytokines, said drug is administered ata dose of about 0.1 to about 100 mg/day.

In the case of cytostatic compounds such as amoxifene, prodasone,sandostatine, polyamine inhibitors or apoptosis inducing compounds suchas sodium butyrate or mitomycin C, said drug is administered at a doseof about 0.1 to about 100 mg/day.

In another advantageous combined preparation of the invention, theimmunotherapy or chemotherapy drug is administered in a repeated way upto 10 times, the interval between each administration being between oneday to two months.

In another advantageous combined preparation of the invention, thechemotherapy or immunotherapy drug and the monocyte derived cells areinjected simultaneously.

In another advantageous combined preparation of the invention, thechemotherapy or immunotherapy drug and the monocyte derived cells areadministered in sequential way, the immunotherapy or chemotherapy drugbeing administered before the monocyte derived cells.

In another advantageous combined preparation of the invention, theinterval of time between the administration of the monocyte derivedcells and the administration of the immunotherapy or chemotherapy drugsis of one day to two months.

In another advantageous combined preparation of the invention, themonocyte derived cells and the chemotherapy or immunotherapy drug areadministered seqentially, the monocytes derived cells being administeredbefore the immunotherapy or chemotherapy drug.

In another advantageous combined preparation of the invention, themonocyte derived cells are administered before the administration of avaccine directed to tumor or infectious antigens, the monocyte derivedcells administration being possibly preceded by a chemotherapytreatment.

In an advantageous embodiment of the invention, the monocyte derivedcells are administered before the administration of the vaccine, thetime internal between the respective administrations being for exampleof one week to three months. The vaccine administration can be repeatedseveral times for optimal immunisation, according to classicalprocedures. In this case, the monocyte derived cells administration canbe considered as a priming for the reaction to the antigen and theadministration(s) of the vaccine as a boost of the immune responses.

In another advantageous embodiment of the invention, the patients arefirst treated by conventional chemotherapy drugs, this treatment beingfollowed sequentially by administration of monocyte derived cells andthen by the vaccine administration as a boost, to induce optimalimmunization against cancer or infectious disease.

In another advantageous combined preparation of the invention, theinterval of time between the administration of the immunotherapy orchemotherapy drug and the administration of the monocyte derived cellsis of one day to two months.

In another advantageous combined preparation of the invention, theadministration of monocyte derived cells is followed by anadministration of chemotherapy or immunotherapy drug.

In another advantageous combined preparation of the invention, theinterval of time between the administration of monocyte derived cellsand the administration of chemotherapy or immunotherapy drugs is of oneday to two months.

The invention also relates to a method for the treatment of residualcancer resistant to chemotherapy or of infectious diseases resistant tochemotherapy comprising the use of a combined preparation of theinvention.

The invention also relates to a method for the treatment of infectiousdiseases resistant to antibiotic treatment comprising the use of acombined preparation of the invention.

The monocytes derived cells which are involved in the invention, can beactivated macrophages and/or monocytes derived antigen presenting cells.

According to an advantageous embodiment, the combined preparation of theinvention comprises monocyte derived cells, loaded with a complexmixture of antigens, and a vaccine containing purified antigens.

The monocyte derived cells loaded with a complex mixture of antigenswill be also referred to as “the cellular vaccine”.

The monocyte derived cells can be macrophages or dendritic cells derivedfrom blood monocytes, preferably cultured in the presence oflymphocytes.

The expression “a complex mixture of antigens” designates antigens witha large spectrum of specificity for the infectious agent or the tumorcells.

As to the vaccine containing purified antigens, its specificity isrestricted to one or a few epitopes of the target agent (infectiousagent or tumor cells).

According to an advantageous embodiment, when the macrophages ordendritic cells derived from blood monocytes are cultured in thepresence of lymphocytes, said lymphocytes are T lymphocytes (CD4⁺ andCD8⁺ types) and natural killer cells (NK cells) generated during thecoculture. These lymphocytes and NK cells can be recovered and possiblyexpanded ex vivo, for simultaneous or sequential injection with saidmonocyte derived cells (loaded with a complex mixture of antigens).

In a particular embodiment, immunomonitoring of patients treated withvaccines containing purified antigens, allows detection andidentification of the specific cellular response to conventionalantigens. The useful specific T cell response can be amplified ex vivo.Monocyte derived cells presenting a mixture of complementary antigenscan then be designed to avoid the development of immunoresistant virusesor tumors.

In the optimal sequence of said cellular vaccine and said vaccinecontaining purified antigens, said vaccine containing purified antigensis used as a boost to achieve immune memory against the targetedinfectious or tumor disease.

The above defined combined preparation of the invention induces cellularand humoral responses to cancer and viral antigens.

According to another advantageous embodiment, the vaccine containingpurified antigens is injected first, followed by the injection of thecellular vaccine.

FIGS. 1 and 2 represent the in vitro synergy between chemotherapy (useof cisplatin) and macrophages (MAK) cytotoxicity on human ovarycarcinoma tumor (IGR-OV1).

FIG. 1 corresponds to chemosensitive tumor and FIG. 2 corresponds tochemoresistant tumor.

Tumor cells have been grown for 3 days of cocultured at 37° C., 5% CO₂from an initial seeding of 10⁵ cells under the following conditions:

presence of cisplatin,

presence of human macrophages,

presence of cisplatin and human macrophages.

The percentage of tumor cell survival is measured according to themethod described in “Feasibility of drug screening with panels of humantumor cell lines using a microculture tetrazolium assay” (Alley M. C.,Scudiero D. A., Monks A., et al.; Cancer Res., 1988, 48: 489-501), andis plotted against the dose of cisplatin (abscissa) in the test tube(molar concentration).

The amount of macrophages used in the experiment is constant.

The initial ratio between macrophage and tumor cells is 4/1 for FIG. 1,and 1/1 in FIG. 2.

The dotted line corresponds to the addition of macrophages alone.

The open circle curve corresponds to the addition of cisplatin alone.

The dark square curve corresponds to the addition of macrophages andcisplatin.

The open triangle curve corresponds to the theoretical addition of theeffects of macrophages alone, plus cisplatin alone.

On FIG. 2, the open circle curve and the open triangle curve aresuperimposed.

Additive effects of macrophages and cisplatin are seen on chemosensitivetumor cells. Synergy or potentiation of macrophages and cisplatin isobserved for the chemoresistant tumor.

FIG. 3 represents the percentage of cellular survival as a function ofcisplatin concentration (M).

The curve with hollow circles corresponds to the use of cisplatin alone.

The plain curve corresponds to the use of MAK alone.

The curve with black squares corresponds to the use of the combinationof MAK followed by cisplatin.

The curve with hollow triangles corresponds to the theoretical additivecurve of a treatment with MAK alone and a treatment with cisplatinalone.

EXAMPLES

The following examples describe some applications of the invention:

1) The synergy between macrophage immunotherapy and chemotherapy hasbeen demonstrated in vitro on a carcinoma tumor cell line. Relativesensitivity of a human ovary tumor cell line and a derived lineresistant to cisplatinum is documented, as well as the cytotoxicity ofactivated macrophages on these lines. Additive antitumoral effects formacrophages and cisplatinum is documented, allowing an effective doseresponse with lower levels of the drug, as demonstrated in FIG. 1 andFIG. 2.

16R-OV1/DDP human ovary cancer cell line was rendered resistant tocisplatinum by continuous exposure to increasing concentrations ofcisplatin (Fajac A., et al. Cisplatin induced apoptosis and p.53 genestatus in a cisplatin resistant human ovarian carcinoma cell line. Int.J. Cancer 68, 67-74, 1996). The cells are grown at 37° C., 5% CO₂ inRPMI 1640 medium in the presence of activated human macrophages (MAK)during 24 hours and then with cisplatin at increasing concentrations.Cisplatin alone (0) inhibited survival by 35% at maximal dose (10⁻³M,see FIG. 3). Activated macrophages inhibited survival by 10%(effector/tumor ratio 4, basal line). The combination of MAK and of alow dose cisplatin (10⁻⁶M) inhibited survival by 40% (). The sequentialcombination of MAK and cisplatin (MAK followed by cisplatin) acted insynergy since the cytotoxicity was much higher at low cisplatin dosethan the theoretical additive curve of both treatments (Δ, see FIG. 3).

2) Nude mice inoculated with human carcinoma solid tumors are initiallytreated with cytotoxic drugs (Adriamycin, Etretinate, Taxotere), usedalone or in combination. After a first response, the tumors grow againand the animals are treated systemically, or locally by injection of 1million activated human macrophages which allow tumor stabilization. Asecond treatment with the same cytotoxic drugs used initially allowfurther antitumoral effect documented by measurement of subcutaneoustumor size.

3) Three patients with colorectal cancer and four patients with lungmesothelioma became resistant to 5-fluorouracil+Cisplatin (or itsoxaliplatin derivative) chemotherapy. They then have been injected withautologous activated macrophages and they have presented tumorstabilization or partial response illustrated by radiography. After afew months, the tumor relapsed and cancer evolution was reported. Asecond chemotherapy treatment, with similar cocktail of cytotoxic drugs,induced complete responses or major partial responses. This indicates amodification of the chemoresistance caused by immunotherapy.

4) Patients with prostate cancer treated by radio and chemotherapypresent a 50% relapse rate of their cancer within 2 years. A treatmentwith activated macrophages is proposed after the conventional therapy.The time of relapse within 2 years as well as the evolution of the tumorare documented.

5) Bacterial infections induced in nude mice are relatively resistant toantibiotics. Effective therapy is achieved by sequential injection ofmacrophages and of antibiotics at usually ineffective doses. Theadditive effects of classical anti-infections drugs and of macrophageimmunotherapy are documented.

6) Patients with myeloid leukemia, or with multiple myeloma, are treatedwith high dose chemotherapy. During the 6 weeks of aplasia, they presentmultiple infections, in particular nosocomial infections. Injections ofactivated macrophages during this period is performed to preventinfections and to allow a cure at lower doses of antibiotics.

7) C57B16 mice bearing solid carcinoma are injected intraperitonealywith a drug inducing apoptosis (1 mg mitomycin C or 0.1 mg sodiumbutyrate). After 24 h, mice are injected with 0.1 million monocytederived cells in tumor periphery. Tumor regression and protectionagainst further tumor challenge is observed only after this combinedtreatment. In another protocol, carcinoma cells are treated in vitrowith 0.01 mM sodium butyrate, and then submitted to phagocytosis bymurine monocyte derived cells. Injection of mice with 0.1 million ofthese monocyte derived cells protects the animals against carcinomachallenge.

8) Dendritic cells (DC) are obtained from bone marrow precursors ofBalb/c syngeneic mice after 7 days of culture in medium supplementedwith GM-CSF and IL-13. The dendritic cells are loaded with the S proteinof the hepatitis B virus at 20 μg/ml for 4 hours. One million cells areinjected intravenously. 7 days later, a mixture of HBS protein andadjuvant is injected in the peritoneal cavity. At day 15, the immuneresponse is assessed by two different methods:

serum titer of antibody against HBS is measured by ELISA,

the spleen from the mice is removed and T lymphocytes are stimulatedwith irradiated isogenic splenocytes loaded with peptides for 7 days. Atday 22, the significant cytotoxic activity of the T lymphocytes ismeasured using as targets p815 cells loaded with the 28-39 peptide (animmunogenic peptide of the S antigen).

9) Patients, whose primary melanoma tumor was removed by surgery, aretreated with chemotherapy agent (DTIC) (dacarbazine) after relapse. Whentheir blood count is back to normal, blood is drawn up through apheresisin order to prepare large amounts of MD-APCs. These cells are thenincubated for 4 hours with allogeneic tumor extract. 3 weeklysub-cutaneous injections (at 4 different sites) of 10⁷ cells are made.Two months later, a cocktail of three antigens (MAGE-3, MELAN A andgp-100) (Boon et al. Immunology today, June 1997, Vol. 18, n 6, 267)plus adjuvant is injected to the patients in order to boost the immunesystem. The increased immune response is monitored by measuring thenumber of antigen specific CD8 T lymphocytes by ELISPOT technique (Herr.et al. Detection and quantification of blood-derived CD8+ T lymphocytessecreting tumor necrosis factor alpha in response to HLA-A2.1-bindingmelanoma and viral peptide antigens. J Immunol Methods 191, no.2:131-42.) It is also assessed that the relapse-free time issignificantly increased.

In a particular embodiment of the invention, macrophages are loaded exvivo with a drug as promyxin (a bioreductive agent) active in hypoxicareas. In this case, the macrophages having been fed with the drug,concentrate in the necrotic/hypoxic area, kill tumor cells in contactand release locally during several days the cytotoxic drug killing theremaining cancer cells. A radiotherapy enhancer (tirazone) is alsoloaded into macrophages which cause, after reinjection, a potentiationof radiotherapy at specific tumor sites.

In another embodiment of the invention, an antibiotic is loaded intomacrophages from patients with nosocomial infections resistant toconventional antibiotics.

The proper sequence and timing of macrophages injections, allowingmaximum activity at the tumor or infectious site, are disclosed.

What is claimed is:
 1. Preparation comprising the following individualcomponents, in the form of a kit: monocyte derived cells which have beenin culture for 5 to 10 days, and chemotherapy drugs, for thesimultaneous, separate or sequential use, for the treatment of cancer orinfectious diseases in a patient.
 2. Preparation according to claim 1,wherein the monocyte derived cells are prepared according to the methodcomprising the following steps: 1) recovering blood derived mononuclearcells directly from blood apheresis or from blood bag collection,followed if necessary by centrifugation, to eliminate the red bloodcells granulocytes and platelets, and collection of peripheral bloodleukocytes; 2) washing peripheral blood leukocytes obtained at thepreceding steps by centrifugation (to remove 90% of platelets, red bloodcells and debris) to obtain mononuclear cells; 3) resuspending the totalmononuclear cells (monocytes+lymphocytes) obtained at the preceding stepin culture medium (RPMI or IMDM type) at 10⁶ to 2.10⁷ cells/ml,completed by at least one of cytokines and autologous serum, and cultureat 37° C. under O₂/CO₂ atmosphere in hydrophobic gas permeable bags, toobtain monocyte derived cells and contaminating lymphocytes. 3.Preparation according to claim 1, wherein the chemotherapy drug isselected from the group of compounds consisting of anthracyclins,daunorubicin, adriamycin, taxoter derivatives, vinca alcaloids,vincristine, taxol, carmustine, cisplatin, fluorouracils, polyamineinhibitors, topoisomerase inhibitors, tamoxifene, prodasone,sandostatine, sodium butyrate, mitomycin C, penicillins, β-lactamines,cephalosporines, cyclines, aminoglucosides, macrolides, sulfamides, AZT,protease inhibitors, acyclovir, retrovir and foscarnet.
 4. Preparationaccording to claim 1, wherein the monocyte derived cells and thechemotherapy drugs are in the form of injectable solutions. 5.Preparation according to claim 4, wherein the form of injectablesolutions are for locally injectable solutions.
 6. Preparation accordingto claim 4, wherein the form of injectable solutions permit systemicallyinjectable solutions.
 7. A process for preparing monocyte derived cellsand chemotherapy drugs in the form of a kit for simultaneous, separate,or sequential use for the treatment of cancer or infectious diseases ina patient, comprising the following steps: 1) recovering blood derivedmononuclear cells directly from blood apheresis or from blood bagcollection, followed if necessary by centrifugation, to eliminate thered blood cells granulocytes and platelets, and collection of peripheralblood leukocytes; 2) washing peripheral blood leukocytes obtained at thepreceding steps by centrifugation (to remove 90% of platelets, red bloodcells and debris) to obtain mononuclear cells; 3) resuspending the totalmononuclear cells (monocytes+lymphocytes) obtained at the preceding stepin culture medium (RPMI or IMDM type) at 10⁶ to 2.10⁷ cells/ml,completed by at least one of cytokines and autologous serum, and cultureat 37° C. under O₂/CO₂ atmosphere in hydrophobic gas permeable bags, toobtain monocyte derived cells and contaminating lymphocytes; and 4)assembling said monocvte derived cells in a kit with chemotherapy drugs.8. The process according to claim 7, further comprising centrifugingsaid monocyte derived cells, washing, and resuspending said monocytederived cells to obtain a suspension of the monocyte derived cells. 9.The process according to claim 8, further comprising the additional stepof freezing said suspension at a temperature below or equal to −80° C.aliquots with the addition of a cryopreservative.
 10. The processaccording to claim 9, further comprising melting said frozen aliquots toobtain a suspension of monocyte derived cells, washing said suspension,and resuspending said suspension in an isotonic medium to obtain asuspension of monocyte derived cells.
 11. Process for the simultaneous,separate, or sequential use of a preparation for the treatment of canceror infectious diseases in a patient, comprising: administering to saidpatient an effective amount of said preparation, wherein saidpreparation comprises the following individual components, in the formof a kit: monocyte derived cells which have been in culture for 5-10days, and chemotherapy drugs.
 12. The process according to claim 11,wherein the monocyte derived cells are administered at a dose from 10⁷to 10¹⁰ monocyte derived cells per injection.
 13. The process accordingto claim 12, wherein the monocyte derived cells are administered at adose from 10⁸ to 10⁹.
 14. The process according to claim 12, wherein themonocyte derived cells are administered repeatedly up to ten times, theinterval between each administration being between three days to twomonths.
 15. The process according to claim 11, wherein the chemotherapydrug is administered at a dose of 0.1 to 1000 mg/day.
 16. The processaccording to claim 11, wherein in the case of administration of a drug,said drug is selected from the group consisting of cytotoxic compounds,cytostatic compounds, compounds inducing apoptosis or cytokines, saiddrug administered at a dose of 0.1 to 100 mg/day.
 17. The processaccording to claim 11, wherein the chemotherapy drug is administeredrepeatedly up to 10 times, the interval between each administrationbeing between one day to two months.
 18. The process according to claim11, wherein the chemotherapy drug and the monocyte derived cells areinjected simultaneously.
 19. The process according to claim 11, whereinthe chemotherapy drug and the monocyte derived cells are administeredsequentially, the chemotherapy drug being administered before themonocyte derived cells.
 20. The process according to claim 19, whereinthe interval of time between the administration of the monocyte derivedcells and the administration of the chemotherapy drugs is of one day totwo months.
 21. The process according to claim 11, wherein the monocytederived cells and the chemotherapy drug are administered sequentially,the monocyte derived cells being administered before the chemotherapydrug.
 22. The process according to claim 11, wherein the monocytederived cells are administered before the administration of a vaccinedirected to tumor or infectious antigens, the monocyte derived cellsadministration being preceded by a chemotherapy treatment.
 23. Theprocess according to claim 22, wherein the interval of time between theadministration of the chemotherapy drug and the administration of themonocyte derived cells is one day to two months.
 24. The processaccording to claim 19, wherein the administration of monocyte derivedcells is followed by an administration of the chemotherapy drug.
 25. Theprocess according to claim 24, wherein the interval of time between theadministration of monocyte derived cells and the administration ofchemotherapy drugs is one day to two months.